LOCUS       62056_19515        4657 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb127, complete sequence.
ACCESSION   MW476665
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4657)
  AUTHORS   Damalas SG, Batianis C, Martin-Pascual M, de Lorenzo V, Martins Dos 
            Santos VAP.
  TITLE     SEVA 3.1: enabling interoperability of DNA assembly among the SEVA, 
            BioBricks and Type IIS restriction enzyme standards
  JOURNAL   Microb Biotechnol 13 (6), 1793-1806 (2020)
  PUBMED    32710525
REFERENCE   2  (bases 1 to 4657)
  AUTHORS   Damalas SG, Batianis C, Martin Pascual M, de Lorenzo V, Martins dos 
            Santos VAP.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JAN-2021) Laboratory of Systems and Synthetic Biology,
            Wageningen University, Stippeneng 4, Wageningen, Gelderland 6708 WE,
            The Netherlands
REFERENCE   3  (bases 1 to 4657)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb 
            Biotechnol"; date: "2020"; volume: "13"; issue: "6"; pages: 
            "1793-1806"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-JAN-2021) Laboratory of Systems and Synthetic Biology, 
            Wageningen University, Stippeneng 4, Wageningen, Gelderland 6708 WE,
            The Netherlands"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      23..57
                     /label=promoter BBa_J23105
                     /note="promoter BBa_J23105"
                     /regulatory_class="promoter"
     regulatory      66..77
                     /label=RBS.20
                     /note="RBS.20"
                     /regulatory_class="ribosome_binding_site"
     CDS             84..797
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     misc_feature    804..824
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      855..949
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1022..1113
                     /label=AmpR promoter
     CDS             1114..1971
                     /label=AmpR
                     /note="beta-lactamase"
     terminator      1978..2017
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            2164..2272
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2336..3481)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     rep_origin      complement(3874..4405)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4532..4618)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gaattcgcgg ccgcttctag agtttacggc tagctcagtc ctaggtacta tgctagctac
       61 tagagtattt aaggctgtac tagatgcgta aaggcgaaga actgttcacg ggcgtagttc
      121 cgattctggt cgagctggac ggcgatgtga acggtcataa gtttagcgtt cgcggtgaag
      181 gtgagggcga cgcgaccaac ggcaaactga ccctgaagtt catctgcacc accggtaaac
      241 tgccggtgcc ttggccgacc ttggtgacga cgttgacgta tggcgtgcag tgttttgcgc
      301 gttatccgga ccacatgaaa caacacgatt tcttcaaatc tgcgatgccg gagggttacg
      361 tccaggagcg taccatttcc ttcaaggatg atggctacta caaaactcgc gcagaggtta
      421 agtttgaagg tgacacgctg gtcaatcgta tcgaattgaa gggtatcgac tttaaagagg
      481 atggtaacat tctgggccat aaactggagt ataacttcaa cagccataat gtttacatta
      541 cggcagacaa gcaaaagaac ggcatcaagg ccaatttcaa gattcgccac aatgttgagg
      601 acggtagcgt ccaactggcc gaccattacc agcagaacac cccaattggt gacggtccgg
      661 ttttgctgcc ggataatcac tatctgagca cccaaagcgt gctgagcaaa gatccgaacg
      721 aaaaacgtga tcacatggtc ctgctggaat ttgtgaccgc tgcgggcatc acccacggta
      781 tggatgaact gtacaaataa taatactagt agcggccgct gcagtccggc aaaaaagggc
      841 aaggtctagt cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
      901 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc
      961 caataattac gatttaaatt agtagcccgc ctaatgagcg ggcttttttt taattcccct
     1021 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1081 taaatgcttc aataatattg aaaaaggaag agtatgagca ttcagcattt tcgtgtggcg
     1141 ctgattccgt tttttgcggc gttttgcctg ccggtgtttg cgcatccgga aaccctggtg
     1201 aaagtgaaag atgcggaaga tcaactgggt gcgcgcgtgg gctatattga actggatctg
     1261 aacagcggca aaattctgga atcttttcgt ccggaagaac gttttccgat gatgagcacc
     1321 tttaaagtgc tgctgtgcgg tgcggttctg agccgtgtgg atgcgggcca ggaacaactg
     1381 ggccgtcgta ttcattatag ccagaacgat ctggtggaat atagcccggt gaccgaaaaa
     1441 catctgaccg atggcatgac cgtgcgtgaa ctgtgcagcg cggcgattac catgagcgat
     1501 aacaccgcgg cgaacctgct gctgacgacc attggcggtc cgaaagaact gaccgcgttt
     1561 ctgcataaca tgggcgatca tgtgacccgt ctggatcgtt gggaaccgga actgaacgaa
     1621 gcgattccga acgatgaacg tgataccacc atgccggcag caatggcgac caccctgcgt
     1681 aaactgctga cgggtgagct gctgaccctg gcaagccgcc agcaactgat tgattggatg
     1741 gaagcggata aagtggcggg tccgctgctg cgtagcgcgc tgccggctgg ctggtttatt
     1801 gcggataaaa gcggtgcggg cgaacgtggc agccgtggca ttattgcggc gctgggcccg
     1861 gatggtaaac cgagccgtat tgtggtgatt tataccaccg gcagccaggc gacgatggat
     1921 gaacgtaacc gtcagattgc ggaaattggc gcgagcctga ttaaacattg gtaaaccgat
     1981 acaattaaag gctccttttg gagccttttt ttttggacga ccccggtcct tttccgctgc
     2041 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     2101 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     2161 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     2221 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgtaggccgg
     2281 ccgcgatgca ggtggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt
     2341 tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact
     2401 cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc
     2461 acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt
     2521 cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt
     2581 agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg
     2641 ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc
     2701 caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg
     2761 ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg
     2821 tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca
     2881 ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt
     2941 tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg
     3001 tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg
     3061 caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg
     3121 cggcctgctt ggcttcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct
     3181 tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct
     3241 gttcgagacg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca
     3301 gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gactatcgag ccgacggact
     3361 ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg
     3421 cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca
     3481 ttcaccctcc ttgcgggatt gccccggaat taattccccg gatcgatccg tcgatcttga
     3541 tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt
     3601 cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg tccataaaac
     3661 cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct
     3721 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     3781 ctgcggactg gctttctacg tggctgccat ttttggggtg aggccgttcg cggccgaggg
     3841 gcgcagcccc tggggggatg ggaggcccgc gttagcgggc cgggagggtt cgagaagggg
     3901 gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg ttaaaaacaa
     3961 ggtttataaa tattggttta aaagcaggtt aaaagacagg ttagcggtgg ccgaaaaacg
     4021 ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga cagcccctca aatgtcaata
     4081 ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt caaggatcgc gcccctcatc
     4141 tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc acttatcccc aggcttgtcc
     4201 acatcatctg tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc gaggctggcc
     4261 agctccacgt cgccggccga aatcgagcct gcccctcatc tgtcaacgcc gcgccgggtg
     4321 agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc aagttttccg
     4381 cgaggtatcc acaacgccgg cggccctaca tggctctgct gtagtgagtg ggttgcgctc
     4441 cggcagcggt cctgatcccc cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4501 ctacggcgcg cccagctgtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg
     4561 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     4621 gcctttaatt aatagctttc gctaaggatg atttctg
//
